Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D273/00—Heterocyclic compounds containing rings having nitrogen and oxygen atoms as the only ring hetero atoms, not provided for by groups C07D261/00 - C07D271/00
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K11/00—Depsipeptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
  • C07K11/02—Depsipeptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof cyclic, e.g. valinomycins ; Derivatives thereof
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides

Definitions

• the present invention relates to novel derivatives having a cyclo-tetradepsipeptide skeletal structure in common to PF 1022, which is a cyclodepsipeptide and has anthelmintic activities, and having excellent anthelmintic activities; and also to anthelmintic agent containing the derivatives.
• the novel PF 1022 derivatives according to the present invention show excellent anthelmintic activities against worms parasitic on animals and therefore are useful as anthelmintic agent.
• the PF 1022 substance is a cyclodepsipeptide which was found as a result of a study on anthelmintic compounds against fowl roundworms see Japanese Patent Application Laid-Open (Kokai) No. HEI 3-35796, European Patent Application Publication No. 0382173A2 and "J. Antibiotics", 45, 692(1992)!.
• the PF 1022 substance is a fermentation product, which is produced by the culture of a filamentous fungus PF1022 strain (deposited under FERM BP-2671 with National Institute of Bioscience and Human-Technology Agency in Tsukuba-shi under the provisions of the Budapest Treaty) belonging to Agonomycetales and is a cyclodepsipeptide represented by the following formula (A): ##STR2##
• the PF 1022 substance is a depsipeptide which is formed of L-N-methylleucine (CH 3 ) 2 --CH--CH 2 --CH(NH--CH 3 )COOH!, D-lactic acid CH 3 --CH(OH)COOH! and D-phenyllactic acid C 6 H 5 --CH 2 --CH(OH)COOH! via ester-bonds and amido-bonds and which can also be represented by the following formula (A'):
• MeLeu is an N-methylleucine residue represented by the following formula: ##STR3## Lac is a lactic acid residue represented by the following formula: ##STR4## and PhLac is a phenyllactic acid residue represented by the following formula: ##STR5##
• the PF 1022B substance of the following formula (B), the PF1022C substance of the formula (C), the PF 1022D substance of the formula (D) and the PF 1022E substance of the formula (E) are produced in addition to the above PF 1022 substance. They have anthelmintic activities and were isolated by the present inventors see Japanese Patent Application No. HEI 3-163085, now Japanese Patent Application Laid-Open (Kokai) No. HEI 5-170749; but concerning PF 1022E, Japanese Patent Application No. HEI 4-279094 (filed Oct. 19, 1992 but not yet laid open)!. ##STR6##
• the PF 1022E substance is a novel compound which has not been reported yet.
• PF 1022 substance and PF 1022B to E substances all possess anthelmintic activities and have a marked structural characteristic in that they have a large cyclo-tetradepsipeptide structure as a basic skeleton, and that they have, as side chains, not only four N-methyl groups and four isobutyl groups but also 0-3 methyl group(s) and 1-4 benzyl group(s) and contain eight asymmetric carbon atoms in their molecules.
• a 24-membered ring is formed via 4 ester bonds and 4 amide-bonds. This structure can be presumed to play an important role on the development of biological activities.
• the PF 1022 substance is a fermentation product of the above-described filamentous fungus.
• Makoto Ohyama et al. proposed, as a process of preparing the PF 1022 substance by total synthesis, a process which comprises the steps shown in the following reaction route map (A) see Japanese Patent Application No. HEI 4-131139 (filed: May 22, 1992) and Japanese Patent Application Laid-Open No. HEI 5-320148 (laid open: Dec. 31 1993)!. ##
• a series of novel derivatives or related products of the PF 1022 substance can each be synthesized either by hydrogenating, in the presence of a rhodium catalyst under mild reaction conditions, one or more phenyl groups in the plural benzyl groups of the PF 1022 substance or the PF 1022B substance or the PF 1022E substance so as to form cyclohexyl group(s), or by chemically modifying the phenyl groups through substitution reaction.
• a series of novel derivatives of the PF 1022 substance can each be prepared in accordance with total synthesis procedures by using L-N-methylleucine (L-MeLeu) or L-leucine in combination with an ⁇ -hydroxycarboxylic acid, particularly a D- or L-lactic acid derivative, which may contain a substituent on its ⁇ -carbon atom, and then condensing the carboxyl group of the leucine compound with the ⁇ -hydroxyl group of the lactic acid compound through an ester bond, condensing the carboxyl group of the resultant esterified product with the amino group of the leucine compound through an amide bond, continuing further condensation of the condensation product as needed, thereby to synthesize a chain-like tetradepsipeptide, followed by cyclizing the tetradepsipeptide.
• L-MeLeu L-N-methylleucine
• L-leucine L-leucine in combination with an ⁇ -hydroxycarboxylic acid, particularly a
• a series of novel PF 1022 derivatives synthesized as described above by the present inventors can generically be represented by the below-described formula (I). It has been ascertained by animal tests that these synthesized novel derivatives have useful anthelmintic activities.
• a cyclodepsipeptide namely a PF 1022 derivative represented by the following formula: ##STR8## wherein (i) R 2 and R 4 are each a cyclohexylmethyl group or benzyl group, R 1 and R 3 are each a methyl group or cyclohexylmethyl group or benzyl group, and X, Y, Z and Q are each a methyl group, provided that at least one of R 1 , R 2 , R 3 and R 4 is a cyclohexylmethyl group, or (ii) R 1 , R 2 , R 3 and R 4 are each a linear or branched alkyl group containing 1 to 11 carbon atoms and may be the same or different from each other, and X, Y, Z and Q are each a methyl group, or (iii) R 1 and R 3 are each a linear or branched alkyl group containing 1 to 11 carbon atoms and may be the same or different from each other, and
• the above-described novel PF 1022 derivative of the formula (I) embraces therein, as preferred embodiments, such hydrogenated derivatives of the PF 1022 substance, as represented by below-described formula (I-i-a); such hydrogenated derivatives of the PF 1022B substance, as represented by the below-described formula (I-i-b); cyclodepsipeptides of the below-described formula (I-ii), cyclodepsipeptides of the below-described formula (I-iii), cyclodepsipeptides of the below-described formula (I-iv), cyclodepsipeptides of the below-described formula (I-v), cyclodepsipeptides of the below-described formula (I-vi-a) and cyclodepsipeptides of the below-described formula (I-vi-b).
• R 1c , R 2c , R 3c and R 4c are each a linear or branched alkyl group containing 1 to 11 carbon atoms, particularly an alkyl group containing 1 to 6 carbon atoms and may be the same or different from each other.
• R 1d and R 3d are each a linear or branched alkyl group containing 1 to 11 carbon atoms, particularly an alkyl group containing 1 to 6 carbon atoms, and may be the same or different from each other, provided that R 1d and R 3d do not stand for methyl groups at the same time.
• R 1e , R 2e and R 3e are each a linear or branched alkyl group containing 1 to 11 carbon atoms, particularly an alkyl group containing 1 to 6 carbon atoms and may be the same or different from each other, and G, L and M denote independently a hydrogen or a substituent, particularly a halo group, hydroxyl group, an alkoxy group, a lower alkenyloxy group, a phenyl-lower alkoxy group, an alkylcarbonyloxy group, tetrahydropyranyloxy group or trityloxy group.
• Cyclodepsipeptides represented by the following formula: ##STR14## wherein at least one of X a , Y a , Z a and Q a is a hydrogen and the reminders thereof are all methyl groups; and preferably either X a and Z a are methyl groups while Y a and Q a are hydrogens, or X a and Z a are hydrogens while Y a and Q a are methyl groups.
• Cyclodepsipeptides represented by the following formula: ##STR15## wherein G',L' and M' denote independently a substituent, particularly a halo group, hydroxyl group, an alkoxy group, a lower alkenyloxy group, a phenyl-lower alkoxy group, an alkylcarbonyloxy group, tetrahydropyranyloxy group or trityloxy group.
• Cyclodepsipeptides represented by the following formula: ##STR16## wherein G', L' and M' denote independently a substituent, particularly a halo group, hydroxyl group, an alkoxy group, a lower alkenyloxy group, a phenyl-lower alkyl group, an alkylcarbonyloxy group, tetrahydropyranyloxy group or trityloxy group.
• Me stands for a methyl group
• Bn a benzyl group
• ChxyMe a cyclohexyl methyl group
• i-Pr an isopropyl group
• n-Bu an n-butyl group
• sec-Bu a secondary butyl group
• i-Bu an isobutyl group
• Example 1 the substances (substance code: PF1022-AHH, -ADH and -BTH) of Example 1 are examples of the derivative represented by the general formula (I-i-a) or (I-i-b), while the substances (substance code: PF1022-209 and -217) of Example 13 and Example 15 are examples of the derivative represented by the general formula (I-ii).
• the substances (substance code: PF1022-203, -205, -207 and -225) of Examples 9, 10, 11 and 12 are examples of the derivative represented by the general formula (I-iii).
• the substance (substance code: PF1022-216) of Example 14 is an example of the derivative represented by the general formula (I-iv).
• the substances (substance code: PF1022-218 and -219) of Examples 16 and 17 are examples of the derivative represented by the general formula (I-v).
• the substances (substance code: PF1022-201 and -202) of Examples 7 and 8 are examples of the derivative represented by the general formula (I-vi-a).
• the substances (substance code: PF1022-005, PF1022E, PF1022-215, -006, -011, -012, -013, -016, -018, -019, -020, -021, -022, -023, -025, -026, -029, -224 and -223) of Examples 5, 6 and 18-34 are examples of the derivative represented by the general formula (I-vi-b).
• hydrogenated derivatives or hydro-derivatives of the general formula (I-i-a) or (I-i-b) can each be synthesized by using, as a starting material, the PF1022 substance or PF1022B substance as prepared by a fermentative method.
• Chemical modification, particularly hydrogenation, of a benzene ring such as a phenyl group is generally regarded to be more difficult than the other reactions such as nitration or acylation which is the reaction by an electrophilic substitution.
• hydrogenation made at high temperature and under high pressure is widely made to reduce a phenyl group into a cyclohexyl group.
• the PF1022 substance has a complex chemical structure as occurred naturally. It is presumed that if the PF 1022 substance is subjected to hydrogenation under conventional conditions at high temperature and under high pressure, decomposition reaction can also be involved by hydrogenolysis.
• Procedures of hydrogenation under milder reaction conditions that is, hydrogenation made at normal temperature under normal pressure are desirable for effecting the reduction of the phenyl groups in the PF 1022 substance. From such a viewpoint, the present inventors have proceeded with an investigation on the usability of various reducing catalysts. As a result, it has been found that a rhodium catalyst is most suitable for the hydrogenation of the phenyl groups in the side-chain benzyl groups of the PF1022 substance to form cyclohexyl groups.
• Examples of reducing catalysts which are usable in the process for preparing the invention derivative of the general formula (I-i-a) or (I-i-b) from the PF1022 substance by hydrogenation, include rhodium, and rhodium-carrier catalysts such as rhodium-carbon and rhodium-alumina, and cationic rhodium complexes such as tris(triphenylphosphine)rhodium. In practice, a rhodium-carbon catalyst is preferred.
• This hydrogenation process is able to minimize the hydrogen pressure and the extent of heating upon the catalytic reduction, but elevated pressure and heating to some extents are allow able so that the reaction time can be shortened and production of by-products can be suppressed.
• an inert solvent such as methanol, ethanol or ethyl acetate
• Isolation of the target product (I-i-a) or (I-i-b) after the reaction can be conducted by a well-known method, for example, filtration, column chromatography or a fractional crystallization method using an inert solvent.
• Derivatives of the general formula (I) according to the present invention can be prepared by a totally synthetic process, that is, by providing the following compounds (1), (2), (3), (4), (5) and (6) and then condensing them successively in proper combinations through an ester-bond or an amide-bond.
• Compound (3) D- or L- ⁇ -hydroxycarboxylic acid, preferably, D- or L-lactic acid or a lactic acid derivative having a desired substituent introduced at its ⁇ -carbon atom, each being represented by the following formula: ##STR18## will hereinafter be abbreviated as A 1 .
• Compound (4) D- or L- ⁇ -hydroxycarboxylic acid represented by the following formula: ##STR19## will hereinafter be abbreviated as A 2 .
• Compound (5): D- or L- ⁇ -hydroxycarboxylic acid represented by the following formula: ##STR20## will hereinafter be abbreviated as A 3 .
• Compound (6): D- or L- ⁇ -hydroxycarboxylic acid represented by the following formula: ##STR21## will hereinafter be abbreviated as A 4 .
• R 1 , R 2 , R 3 and R 4 have the same meanings as R 1 , R 2 , R 3 and R 4 defined in connection with the general formula (I) given hereinbefore.
• the carboxyl group of Compound (1) or (2) is reacted with the ⁇ -hydroxyl group of Compound (3), (4), (5) or (6).
• the following four Compounds (7)-(10) each having an amino group at one end thereof and a carboxyl group at the other end thereof can therefore be prepared as primary condensates containing the ester-linkage.
• the a-hydroxycarboxylic acid which is Compound (4), (5) or (6) can be prepared by reacting a corresponding ⁇ -amino acid with sodium nitrite to convert its amino group into a diazo group (--N 2 ), and then converting the diazo group to a hydroxyl group by acid treatment.
• Such target PF1022 derivative (I) (wherein R 1 ⁇ R 3 , and also R 2 ⁇ R 4 ) can be prepared by using Leucine compound (1) or (2) in combination with Compound (3) or Compound (4), condensing them by esterification, preparing each of intermediates (7) and (8) with forming the amide-bond, producing the chain-like Compound (15) via Compounds (11), and then cyclizing Compound (15), in accordance with the order as illustrated in the below-described reaction route map D. ##STR23##
• R 1 , R 2 , R 3 and R 4 can each be a C 1 -C 11 alkyl group.
• alkyl group examples include methyl, ethyl, propyl, (specifically, n-propyl, iso-propyl), butyl (specifically, n-methyl, iso-butyl, sec-butyl, tert-butyl), pentyl (specifically, n-pentyl, iso-pentyl, sec-pentyl, 1,2-dimethylpropyl, neo-pentyl, 1-ethylpropyl, 1,1-dimethylpropyl), hexyl, heptyl, octyl, nonyl and decyl groups.
• Preferred are lower (C 1 -C 6 ) alkyl groups.
• R 1 , R 2 , R 3 and R 4 each represents a substituted or unsubstituted phenyl or benzyl group
• specific examples of such a group include phenyl; o-, m- and p-hydroxyphenyl; o-, m- and p-(C 1-10 )alkylphenyl; o-, m- and p-(C 1-10 )alkoxyphenyl; o-, m-, and p-halogeno(F, Cl, Br, I)phenyl.
• benzyl examples include benzyl; o-, m- and p-hydroxybenzyl; o-, m- and p-(C 1-10 )alkylbenzyl; o-, m- and p-(C 1-10 )alkoxybenzyl; and o-, m-, and p-halogeno(F, Cl, Br, I)benzyl.
• the number of the substituents on the phenyl nucleus of the benzyl group can be 1-4.
• the amino-protected leucine compound (1) or (2),and ⁇ -carboxyl-protected Compounds (3), (4), (5) and (6), that is, ⁇ -carboxyl-protected ⁇ -hydroxycarboxylic acids are employed.
• a condensation method using the ester-bond it is desired to conduct the condensation with employing Compounds (3)-(6), each of which has its ⁇ -hydroxyl group in the free form, in the presence of a condensation agent.
• the amino-protected leucine compound (1) or (2) can be condensed, through an ester-bond, with a reactive derivative at the ⁇ -OH group of the carboxyl-protected Compound (3), (4), (5) and (6).
• the ⁇ -carboxyl-protected compound (3), (4), (5) or (6) is the D-isomer, it is desired that the compound has been made reactive by substituting the ⁇ -hydroxyl group by a chlorine atom, a bromine atom or the like.
• the compound is the L-isomer, it is desired that the ⁇ -hydroxyl group has been converted into a sulfonate ester such as tosylate, methanesulfonate or the like.
• ester-type compound (7), (8), (9) or (10) in which the carboxyl and amino groups are both protected.
• carboxyl-protecting group examples include those removable under acid hydrolytic or reducing conditions, such as t-butyl, benzyl, p-methoxybenzyl, benzhydryl and trityl groups; and those removable under neutral conditions such as an allyl group.
• amino-protecting group examples include those removable under acid hydrolytic or reducing conditions such as benzyloxycarbonyl, t-butyloxycarbonyl, p-methoxybenzyloxycarbonyl and formyl groups; and those removable under neutral conditions and commonly used in peptide chemistry, such as an aryloxycarbonyl group.
• the protecting group when the protecting group is removable under acid hydrolytic conditions, it is treated with trifluoroacetic acid, methanesulfonic acid, trifluoromethanesulfonic acid or the like. Treatment with trifluoroacetic acid is most preferred.
• treatment under catalytic reduction conditions using a palladium catalyst is desired.
• the protecting group When the protecting group is removable under neutral conditions, for example, an aryl group, aryloxycarbonyl group or the like, it may be reacted with potassium 2-ethylhexanoate in the presence of zero-valence palladium catalyst for the deprotection.
• Compound (11) is prepared, as shown in the reaction route map B, by condensing, the deprotected Compound (7) with the deprotected Compound (8) through an amide-bond.
• Compound (11) obtained by the condensation of Compounds (7) and (8) is condensed further with Compound (9).
• Compound (9) can also be condensed with Compound (10). In this case, it is possible to form an amide-bond between the amino group of Compound (9) and the carboxyl group of Compound (10) or between the amino group of Compound (10) and the carboxyl group of Compound (9).
• Compound (11), which has been obtained by the condensation of Compounds (7) and (8), can be condensed with Compound (14), which has been obtained by the condensation of Compound (9) and Compound (10). In this case, it is possible to form an amide-bond between the amino group of Compound (11) and the carboxyl group of Compound (14) or between the amino group of Compound (14) and the carboxyl group of Compound (11).
• Compound (15) can be prepared by bonding two molecules of Compound (11) together through an amide-bond.
• a derivative of the general formula (I) can be prepared.
• This ring closure is effected by treating Compound (13) or Compound (15) using dicyclohexylcarbodiimide (DCC) or 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI) and an additive N-hydroxysuccinic acid imide (HOSU), 1-hydroxybenzotriazole (HOBt), or the like! in combination.
• DCC dicyclohexylcarbodiimide
• EDCI 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
• HOSU N-hydroxysuccinic acid imide
• HBt 1-hydroxybenzotriazole
• solvents usable in the above ring-closure reaction include ether-solvents such as ether, tetrahydrofuran (THF) and 1,4-dioxane, and aprotic solvents such as N,N-dimethylformamide (DMF), acetonitrile and chloroform.
• ether-solvents such as ether, tetrahydrofuran (THF) and 1,4-dioxane
• aprotic solvents such as N,N-dimethylformamide (DMF), acetonitrile and chloroform.
• DMF N,N-dimethylformamide
• acetonitrile acetonitrile
• chloroform acetonitrile
• the ring-closure reaction can be carried out at 0°-50° C., preferably 20°-30° C.
• the derivatives of the formulas (I-vi-a) and (I-vi-b) can be prepared by introducing, in accordance with known chemical methods, various substituents for the hydrogen(s) on the benzene ring (phenyl group) of the benzyl group, that is, a side chain of the PF1022 substance or in the phenolic hydroxyl group on the p-hydroxyphenylmethyl group (namely, benzyl group), that is, a side chain of the PF1022 E substance refer to Japanese Patent Application HEI 4-279094 (not yet laid open), and the synthetic Example which will be described subsequently in Example 6)!.
• substituents which may be introduced in the benzene ring, namely, the phenyl nucleus of the benzyl group of the PF1022 substance or in the phenolic hydroxyl group on the benzyl group of the PF1022E substance, include linear or branched alkyl groups, alkenyl groups, alkynyl groups, substituted or unsubstituted benzyl groups, diphenylmethyl group, triphenylmethyl group and acyl groups. Particularly preferred are alkanoyl, carbamoyl, methoxymethyl, methylthiomethyl and tetrahydropyranyl groups.
• substituent(s) replaceable for the hydrogen(s) on the benzene ring include halogen atoms, as well as such substituents as replaceable for the hydrogen(s) on an aromatic ring by ordinary electrophilic substitution.
• the above substituent-introducing reaction can be performed in an inert solvent by etherification, acylation, carbamoylation or the like.
• the etherification can be conducted by a reaction with diazomethane or diphenyldiazomethane, a reaction with isobutene or dihydropyrane in the presence of an acid catalyst, or a reaction with an alkyl halide, an alkenyl halide, an alkynyl halide, a benzyl halide, a substituted benzyl halide, or a triphenylmethyl chloride (namely, trityl chloride).
• the acylation can be conducted by a reaction with an acyl halide or alkyl chlorocarbonate in the presence of an organic base such as triethylamine or an inorganic base such as potassium carbonate.
• an organic base such as triethylamine or an inorganic base such as potassium carbonate.
• Most of the well-known reactions for the modification of a phenolic hydroxyl group can be applied to as such.
• the halogenation or other well-known electrophilic substitutions can also be applied.
• Derivatives of the general formula (I) according to the present invention namely, derivatives of the formulae (I-i-a), (I-i-b), (I-ii), (I-iii), (I-iv), (I-v), (I-vi-a) and (I-vi-b) all have useful anthelmintic activities and show a low acute toxicity to mammarian animals.
• novel PF1022 derivatives according to the present invention can be converted to their acid addition salts by reacting them with a pharmaceutically acceptable inorganic acid such as hydrochloric acid, sulfuric acid or phosphoric acid, or a pharmaceutically-acceptable organic acid such as acetic acid, propionic acid, citric acid or methanesulfonic acid.
• a pharmaceutically acceptable inorganic acid such as hydrochloric acid, sulfuric acid or phosphoric acid
• a pharmaceutically-acceptable organic acid such as acetic acid, propionic acid, citric acid or methanesulfonic acid.
• the PF1022 derivatives of the present invention or their salts can be formulated into anthelmintic compositions by mixing them with a pharmaceutically acceptable, solid or liquid carrier.
• an anthelmintic composition characterized in that the composition comprises a novel cyclodepsipeptide represented by the general formula (1) or its salt as an active ingredient.
• the novel derivative of the general formula (I) according to the present invention or the composition containing the derivative can be administered to animals orally or parenterally, for example, rectally.
• the dose of the derivative can be determined depending on the kind of a parasite to be eliminated, the kind of a host animal to be treated and various other factors.
• oral administration of the compound of the formula (I) at a dose of 0.05 mg/kg or greater, preferably 0.2 mg to 3 mg/kg is recognized to exhibit the anthelmintic action against parasites.
• the compound of the general formula (I) of the present invention can be formulated into an anthelmintic composition just in the same manner as for the PF1022 substance, which is described in Japanese Patent Application Laid-Open No. HEI 3-35796 or European Patent Application Publication No. 0382173 A2.
• Examples of the animal to which the PF1022 derivative of the formula (I) of this invention can be applied as an anthelmintic may include domestic animals, poultry, experimental animals and pets, such as swine, cattle, horses, rabbits, sheep, goats, domestic fowls, ducks, turkeys, mice, white rats, guinea pigs, monkeys, dogs, cats and small birds.
• Illustrative parasites on these animals include parasites on cattle and sheep such as twisted stomachworms, stomachworms belonging to the genus Ostertagia, small hairworms, nematodes belonging to the genus Cooperia, nodularworms belonging to the genus Oesophagostomum, amphisomes, intestinal tapeworms (Moniezia benedeni), lung worms and liver flukes; parasites on swine such as roundworms, whipworms and nodularworms; parasites on dogs such as roundworms, hookworms, whipworms and heart worms; parasites on cats such as roundworms and Spirometra mansoni; and parasites of chickens such as roundworms, hairworms and cecal worms.
• parasites on cattle and sheep such as twisted stomachworms, stomachworms belonging to the genus Ostertagia, small hairworms, nematodes belonging to the genus Cooperia, nodularworm
• the compound of the present invention is also effective for the elimination of parasites on human bodies such as roundworms, pinworms, hookworms (Ancylostoma duodenale, Ancylostoma ceylanicum, Necator americanus), oriental hairworms, strongyloides worms and whipworms.
• the novel PF1022 derivative according to the present invention can be used for the treatment and prevention of parasitic infections.
• the derivative may be administered orally or parenterally.
• a liquid preparation of the derivative may be forcedly administered using a stomach catheter or the like, or administered after mixing it with daily feed or drinking water, or administered in an ordinary dosage form suitable for oral administration, such as tablets, capsules, pellets, boluses, powders or soft capsules.
• parenteral administration Upon parenteral administration, it may be administered subcutaneously, intramuscularly, intravenously, intraperitoneally or through a similar route by injecting the derivative of the formula (I) prepared in the form of a water-insoluble preparation in peanut oil, soybean oil or the like or in the form of a water-soluble preparation in glycerol, polyethylene glycol, etc.
• PF1022 derivative for the prevention of parasitic infections, it is a common practice to administer the PF1022 derivative orally as a a mixture of it with daily feed. Although no limitation is imposed on the administration period in the case of preventive purposes, it is, in most cases, sufficient to administer it for about 2 months in the case of broiler chickens and for about 5 months in the case of swine.
• the dose of the PF1022 derivative according to the present invention may vary depending on the kind of the animal to be treated, the kind of the parasite and the method of administration. For instance, when fowl roundworms are to be eliminated by oral administration of a liquid preparation using a stomach catheter, it can be administered at 0.05 mg/kg or more.
• the derivative can be mixed with feed at a concentration of 1 ppm or higher, preferably 5 to 10 ppm and administered continuously.
• a solution or suspension of the PF1022 derivative of the present invention in a liquid carrier can be administered to animals by subcutaneous or intramuscular injection, etc.
• non-aqueous formulations using a vegetable oil such as peanut oil or soybean oil are employed.
• Aqueous parenteral formulations which contain a water-soluble carrier such as glycerol or polyethylene glycol can also be employed for parenteral administration.
• These formulations generally contain the compound of the present invention in an amount of 0.1 to 10 wt. %. Even when the PF1022 derivative of the present invention is orally administered to mice at the dose of 300 mg/kg, normal body-weight gains are obtained without any abnormalities. This indicates the low toxicity of this substance.
• Fowls (three fowls per group), which had been artificially infected with fowl roundworms and whose infection therewith had been confirmed by scatoscopy, were used as experimental animals.
• the test substance weighed in a dose (mg) accurately calculated on the basis of the body weight (kg) of each fowl was suspended in carboxymethylcellulose-containing water, and the resulting suspension was administered orally as a single dose unit using a stomach tube. After the administration, the worms eliminated from the fowls were counted daily. Seven days after the administration, the fowl was sacrificed and autopsied and the worms remaining in its intestinal tract were counted. The percent elimination was calculated in accordance with the following calculation equation: ##EQU1##
• Test results are summarized in Table 2 shown below. Each test substance is indicated by the corresponding substance code name shown above in Table 1.
• T Trichostrongylus colubriformis
• T Haemonchus contortus
• the number of parasite's eggs excreted with feces from the sheep was counted quantitatively before and after the administration so that the degree of anthelmintic effects was determined.
• the anthelmintic effects are evaluated by rated numeral 0, 1, 2 or 3.
• the anthelmintic effects are rated "0" when there were no anthelmintic activities, "2" when excretion of parasite's eggs was observed, and "3" when excretion of parasite's eggs stopped, that is, parasitic worms were removed completely.
• Table 3 shows the results of the test on anthelmintic activities against the above-described two types sheep parasitic worms.
• Anthelmintic effects of certain PF1022 derivatives on a rat intestinal nematode were tested in accordance with the following method.
• PF1022, PF1022E, PF1022-002, PF1022-003, PF1022-209, PF1022-218 and PF1022-219 were, as test substances, forcedly administered p.o. to the rats in the groups, respectively, in an amount of 10 mg/kg per rat.
• each test substance (8 mg) was dissolved in 0.2 ml of dimethyl sulfoxide and then, the resultant solution was diluted with distilled water to give a 2 ml suspension.
• the rats were each subjected to autopsy and imaginal worms parasitic on the small intestines were counted.
• Tr triphenylmethyl group (trityl group)
• VALA 2-hydroxyisovaleric acid residue ##STR34##
• OctA 2-hydroxyoctanoic acid residue ##STR35##
• HOBt 1-hydroxybenzotriazole
• BOP-Cl N,N-bis(2-oxo-3-oxazolidinyl)phosphinic acid chloride
• Fraction 4 (1.5 l): A mixture of partial hydrogenation products of PF1022 and the starting material PF1022 was contained therein.
• PF1022-AHH hexahydro-PF1022
• substance code: PF1022-AHH a compound of the general the formula (I-i-a) where a benzyl group is present as R 2a and a cyclohexylmethyl group as R 4a .
• the resulting reaction mixture was added with 150 ml of ethyl acetate, followed by washing with 80 ml of water, 80 ml of a saturated aqueous solution of sodium bicarbonate, 80 ml of a 5% aqueous solution of potassium hydrogen sulfate and 80 ml of a saturated aqueous solution of sodium chloride and drying over sodium sulfate.
• the solvent was removed from the dried solution.
• PF1022 E substance which can also be expressed as "cyclo-(-L-MeLeu-D-Lac-L-MeLeu-D-PhLac-L-MeLeu-D-Lac-L-MeLeu-D-TYRA)", were dissolved, followed by ice cooling. Under a nitrogen stream, the resulting solution was added with 0.02 ml (0.32 mmol) of methyl iodide and 9 mg (60% in oil, 0.23 mmol) of sodium hydride were added, followed by reaction for 40 minutes (for O-methylation).
• PF1022 E which may also be expressed as "cyclo-(-L-Meleu-D-Lac-L-MeLeu-D-TYRA-L-MeLeu-D-Lac-L-MeLeu-D-PhLac)"
• the resulting reaction mixture was added with 200 ml of ethyl acetate.
• the resulting mixture was allowed to separate into two layers.
• the water layer so obtained was extracted again with ethyl acetate.
• the ethyl acetate extract was combined with the organic layer, followed by washing twice with 50 ml portions of a 30% aqueous solution of sodium chloride, drying over anhydrous magnesium sulfate and then concentrating under reduced pressure, whereby 1.33 g of the title compound were obtained (yield: 24.3%).
• the resulting solution was washed successively with a 7% aqueous solution of sodium bicarbonate, water and a 30% aqueous solution of sodium chloride, each, in an amount of 50 ml, dried over anhydrous magnesium sulfate and concentrated under reduced pressure, whereby 2.68 g of the title compound were obtained (94.4%).
• the product so obtained was fed to as such for use in the next reaction.
• the acidified solution was washed successively with water and a 30% aqueous solution of sodium chloride, dried over anhydrous sodium sulfate and then concentrated, whereby the title compound were obtained.
• the product so obtained was fed to as such, for use in the next reaction.
• the water layer so obtained was extracted four times with 100 ml-portions of ethyl acetate.
• the combined organic layers were dried over anhydrous magnesium sulfate.
• the magnesium sulfate was filtered off and the filtrate was concentrated under reduced pressure, whereby the title compound was obtained.
• Ethyl acetate (300 ml) was added to the compound obtained in the procedure a) to dissolve the latter in the former.
• a solution of diphenyldiazomethane in ethyl acetate (38.8 g/400 ml) was added dropwise, followed by stirring overnight at room temperature.
• Acetic acid (30 ml) was added to the reaction mixture, followed by washing thrice with 300-ml portions of a saturated aqueous solution of sodium bicarbonate. The organic layer was dried over anhydrous magnesium sulfate, followed by filtration.
• the water layer was extracted four times with 100 ml portions of ethyl acetate.
• the combined organic layers were dried over anhydrous magnesium sulfate. After the magnesium sulfate was filtered off, the filtrate was concentrated under reduced pressure. The residue was supplied to as such for use in the next reaction.
• the purified product was lyophilized, whereby 134 mg of the title compound were obtained as white powder (yield: 29.0%).
• the catalyst was filtered off and then, the filtrate was concentrated, whereby 1.21 g of the title compound were obtained as white powder.
• the compound was provided without purification for use in the next reaction.
• the reaction mixture was concentrated and the residue obtained was dissolved in 120 ml of ethyl acetate.
• the resulting solution was washed successively with a 7% aqueous solution of sodium bicarbonate, a 5% aqueous solution of potassium bisulfate and a 20% aqueous solution of sodium chloride, each 120 ml, dried over anhydrous magnesium sulfate and then concentrated under reduced pressure.
• the resulting solution was washed with 80 ml of a 7% aqueous solution of sodium bicarbonate and a 30% aqueous solution of sodium chloride, dried over anhydrous magnesium sulfate and then concentrated, whereby the title compound was obtained as a colorless oil.
• the compound was provided without purification for use in the next reaction.
• the resulting mixture was washed with 65 ml of a 7% aqueous solution of sodium bicarbonate and 35 ml of a 30% aqueous solution of sodium chloride, dried over anhydrous magnesium sulfate and then concentrated, whereby the title compound was obtained as white prism crystals.
• the compound was provided without purification for use in the next reaction.
• the reaction mixture obtained was concentrated and the residue was dissolved in 100 ml of ethyl acetate.
• the resulting solution was washed successively with 100 ml of a 7% aqueous solution of sodium bicarbonate, 100 ml of a 5% aqueous solution of potassium bisulfate and 100 ml of a 30% aqueous solution of sodium chloride, dried over anhydrous magnesium sulfate and concentrated.
• the resulting solution was washed successively with 75 ml of a 7% aqueous solution of sodium bicarbonate and 75 ml of a 30% aqueous solution of sodium chloride, dried over anhydrous magnesium sulfate and concentrated, whereby the title compound was obtained as a colorless oil.
• the compound was provided without purification for use in the next reaction.
• the resulting mixture was washed successively with a 5% solution of sodium bisulfate, a saturated aqueous solution of sodium bicarbonate and a saturated aqueous solution of sodium chloride, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated. Ethyl acetate (100 ml) was added to the concentrate. The resulting mixture was washed successively with a 5% solution of sodium bisulfate, a saturated aqueous solution of sodium bicarbonate and a saturated aqueous solution of sodium chloride, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated.
• the residue was dissolved in 70 ml of ethyl acetate, followed by washing successively with 70 ml of a 5% aqueous solution of potassium bisulfate and 70 ml of a 30% aqueous solution of sodium chloride, drying over anhydrous magnesium sulfate and concentration.
• the reaction mixture was diluted with 40 ml of ethyl acetate and 20 ml of hexane, followed by washing successively with 40 ml of water, 40 ml of a saturated aqueous solution of sodium bicarbonate and 40 ml of a saturated aqueous solution of sodium chloride, filtration using a small quantity of silica gel for chromatography, and drying over anhydrous magnesium sulfate.
• the solvent was distilled off under reduced pressure.
• the water layer was extracted with a liquid mixture of 20 ml of ethyl acetate and 8 ml of hexane.
• the combined organic layers were filtered using a small quantity of silica gel for chromatography, followed by drying over anhydrous sodium sulfate. The solvents were then distilled off under reduced pressure.
• the reaction mixture was diluted with ethyl acetate, followed by washing once with 50 ml of a 5% aqueous solution of potassium bisulfate and twice with 50 ml portions of a saturated aqueous solution of sodium bicarbonate, filtering through a small quantity of silica gel for chromatography, and drying over anhydrous sodium sulfate.
• the solvent was then distilled off under reduced pressure.
• the PF 1022 derivatives represented by the general formula (I) which are provided herein by the present invention each has anthelmintic activities against various parasitic worms which are parasitic on human bodies, domestic animals and companion animals. They are therefore useful as anthelmintic agent for prevention or treatment of parasitic infections.

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• 1994
  • 1994-02-18 KR KR1019950703498A patent/KR100309091B1/ko not_active IP Right Cessation
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